pAAVLINKv1-pCALM1-Cre-3'-CLTC-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-CLTC-FLAG                6874 bp DNA     circular SYN 18-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6874)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    12..130
                     /label=AAV2 ITR
     promoter        145..265
                     /label=pCALM1
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          1309..1349
                     /label=Splicing Donor
     protein_bind    1356..1389
                     /label=lox2272-JTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          1390..1481
                     /label=Splicing Acceptor
                     /note="Splicing acceptor"
     CDS             1482..3587
                     /codon_start=1
                     /label=GLI3
                     /translation="VVQTALSETQDPEEVSVTVKAFMTADLPNELIELLEKIVLDNSVF
                     SEHRNLQNLLILTAIKADRTRVMEYINRLDNYDAPDIANIAISNELFEEAFAIFRKFDV
                     NTSAVQVLIEHIGNLDRAYEFAERCNEPAVWSQLAKAQLQKGMVKEAIDSYIKADDPSS
                     YMEVVQAANTSGNWEELVKYLQMARKKARESYVETELIFALAKTNRLAELEEFINGPNN
                     AHIQQVGDRCYDEKMYDAAKLLYNNVSNFGRLASTLVHLGEYQAAVDGARKANSTRTWK
                     EVCFACVDGKEFRLAQMCGLHIVVHADELEELINYYQDRGYFEELITMLEAALGLERAH
                     MGMFTELAILYSKFKPQKMREHLELFWSRVNIPKVLRAAEQAHLWAELVFLYDKYEEYD
                     NAIITMMNHPTDAWKEGQFKDIITKVANVELYYRAIQFYLEFKPLLLNDLLMVLSPRLD
                     HTRAVNYFSKVKQLPLVKPYLRSVQNHNNKSVNESLNNLFITEEDYQALRTSIDAYDNF
                     DNISLAQRLEKHELIEFRRIAAYLFKGNNRWKQSVELCKKDSLYKDAMQYASESKDTEL
                     AEELLQWFLQEEKRECFGACLFTCYDLLRPDVVLETAWRHNIMDFAMPYFIQVMKEYLT
                     KVDKLDASESLRKEEEQATETQPIVYGQPQLMLTAGPSVAVPPQAPFGYGYTAPPYGQP
                     QPGFGYSM"
     RBS             1639..1647
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    1640..1645
                     /label=SDS
     misc_feature    1966..1971
                     /label=SDS
     CDS             3594..3659
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3663..4094
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3807..3812
                     /label=SDS
     repeat_region   4137..4277
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4137..4266
                     /label=AAV2 ITR
     rep_origin      4352..4807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5089..5193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5194..6054
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6225..6813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggttgtacaa acagctttgt
     1501 ctgagactca ggaccctgaa gaagtgtcag taactgtaaa ggctttcatg actgcagacc
     1561 ttcctaatga actcattgaa ctgctggaga aaattgtcct tgataactct gtattcagtg
     1621 aacacaggaa tctgcaaaac ctccttatcc tcactgcaat taaggctgac cgtacacgtg
     1681 ttatggagta tattaaccgc ctggataatt atgatgcccc agatattgcc aatatcgcca
     1741 tcagcaatga gctgtttgaa gaagcatttg ccattttccg gaaatttgat gtcaatactt
     1801 cagcagttca ggtcttaatt gagcatattg gaaacttgga tcgggcatat gagtttgctg
     1861 aacgttgcaa tgaacctgcg gtctggagtc aacttgcaaa agcccagttg cagaaaggaa
     1921 tggtgaaaga agccattgat tcttatatca aagcagatga tccttcctcc tacatggaag
     1981 ttgttcaggc tgccaatact agtggaaact gggaagaact ggtgaagtac ttgcagatgg
     2041 cccgtaagaa ggctcgagag tcctatgtgg agacagaact gatattcgca ctggctaaaa
     2101 caaaccgcct tgcagagtta gaagaattta tcaatggacc aaataatgct catatccaac
     2161 aagttggtga ccgttgttat gatgaaaaaa tgtatgatgc tgctaagttg ttgtacaata
     2221 atgtttccaa ttttggacgt ttggcatcta ccctggttca cctgggtgaa tatcaggcag
     2281 ctgttgatgg ggctaggaaa gctaacagta ctcgaacatg gaaagaggtc tgcttcgcct
     2341 gtgtagatgg gaaagaattc cgtcttgctc agatgtgtgg acttcatatt gttgtacatg
     2401 cagatgaatt agaagaactt atcaactact atcaggatcg tggctatttt gaagagctga
     2461 tcaccatgtt ggaagcagca ctgggacttg agcgagctca catgggaatg tttactgaat
     2521 tagctattct atactctaaa tttaagcctc agaaaatgag ggagcacctg gagctgttct
     2581 ggtctagagt gaatattccc aaggtgctaa gagctgcaga acaagctcat ctttgggcag
     2641 aactggtgtt tttgtatgac aagtatgaag aatatgataa tgccataatt accatgatga
     2701 atcatccaac tgatgcctgg aaagaagggc aattcaaaga tatcattacc aaggttgcca
     2761 atgtggaact atactacaga gcaatacagt tctacttaga attcaagcct ctgttgttaa
     2821 atgatttgct gatggtgctg tctccacggt tggatcacac tcgtgcagtc aattatttca
     2881 gcaaggttaa acagctacca ctggtgaaac cgtatttgcg ttcagttcag aaccataaca
     2941 acaaatctgt gaatgaatca ttgaacaatc tttttattac agaagaagat tatcaggctc
     3001 tgcgaacatc aatagatgct tatgacaact ttgacaatat ctcgcttgct cagcgtttgg
     3061 aaaaacatga actcattgag ttcaggagaa ttgctgctta tctcttcaaa ggcaacaatc
     3121 gctggaaaca gagtgtagag ctgtgcaaga aagacagcct ttacaaggat gcaatgcagt
     3181 atgcttctga atctaaagat actgaattgg ctgaagaact cctgcagtgg tttttgcagg
     3241 aagaaaaaag agagtgcttt ggagcttgtc tgtttacctg ttacgatctt ttaaggccag
     3301 atgtcgtcct agaaactgca tggaggcaca atatcatgga ttttgccatg ccctatttca
     3361 tccaggtcat gaaggagtac ttgacaaagg tggataaatt agatgcttca gaatcactga
     3421 gaaaagaaga agaacaagct acagagacac aacccattgt ttatggtcag ccccagttga
     3481 tgctgacagc aggacccagt gttgccgtcc ctccccaggc accttttggt tatggttata
     3541 ccgcaccacc gtatggacag ccacagcctg gctttgggta cagcatgtgt acagactaca
     3601 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3661 aactcgagat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3721 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3781 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     3841 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3901 tgacaattcc gtggtgttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3961 ctagctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4021 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     4081 gggaggtttt ttaaagatct ggagaccggt gtggcacgtg cggaccgagc ggccgcagga
     4141 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4201 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4261 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4321 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4381 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     4441 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4501 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4561 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4621 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4681 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4741 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4801 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4861 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4921 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4981 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5041 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5101 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5161 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5221 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5281 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5341 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5401 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5461 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5521 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5581 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5641 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5701 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5761 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5821 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5881 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5941 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6001 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6061 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6121 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6181 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6361 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6601 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6721 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6781 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6841 ttcctggcct tttgctggcc ttttgctcac atgt
//